... as their orientation relative to the gRNA (5’ or 3’). In some cases, there is no requirement, which is... spCas9 isn’t always the winner!).   Fig. 3 – Comparison of different Cas enzymes using CasPEDIA...you are a structure junky who just wants to learn about Cas domains. CasPEDIA either directly has this information...

...very little bit about hiring when we need to do it quickly. Hiring is hard but as a 3 year Boston Globe...like The Atlantic. We’ve been in documentaries in 3 languages (Japanese, German, and English) and even...keep spreading the word about materials sharing, but we don’t have to worry about getting support from depositors...of course always being vigilant about resources, I’m not worried about the stewardship of this in the ...Addgene is about half of what is standard in most sectors. The Addgenies support a high collaboration, high...on this. Addgene’s success is about the Addgenies. I never worry about their dedication, excellent intentions...more about Addgene Resources on the Addgene website Find plasmids for your research  Learn about our ...

...microenvironments made of?  A tumor’s genesis is divided into 3 phases: initiation, progression, and metastasis. The...microenvironment’s physiology is deeply connected to these 3 stages and contains multiple cell types: fibroblasts... related blog posts Learn about our cancer collection pages Read more about the tumor microenvironment...the tumor. How do microenvironment enzymes and metabolic reprograming influence tumors development? Being... vice versa, let’s take a look at cancer cell metabolism. Cancer cells promote oxidative stress leading...of cells affect tumor progression, enzymes and metabolism reprogramming aren’t the whole story. It’s essential...particularly interested in science journalism and writing about biomedical issues. Follow her on Twitter @BrbaraPinho6...

...way is not necessarily easy, so we came up with a 3-part strategy: Train: Nearly every new Addgene employee...communication has helped Addgene move from completing 2-3 large projects a year, on average, to completing 8...differently based on their audience. How you talk about your experiments to your PI, your colleagues, or...have cross-team projects within your own lab, collaborations with other scientists within and outside your...not happen in the lab, they have parallels to collaborative lab work. Corralling context and junking jargon...difficult to know what kind of plate they were talking about during the meetings. And “plate” certainly wasn’... a lot on context. New people in the lab or collaborators will not have the context you do, which means...

...they observed that 41/42 embryos were non-mosaic at 3 days post fertilization. Since Ma et al. designed ...proper development. Ma et al.’s experiments showed a 3% mosaic rate, and this rate would need to be much,...August 2nd in Nature gives us even more to talk about, as it represents another step towards CRISPR germline...impossible with current tools, thus quieting some fears about the creation of “designer babies.” The differences...of individuals with varied backgrounds. Talking about this study with non-biologist friends Like me, you... Blog Check out Our New CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes...

...workflow. B) Detail of spacer peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm single alpha helix) and target...optimized PASSIGE plasmids here!     Figure 3: Schematic of the PASSIGE strategy. First, twin prime.... When the prime editor releases the DNA, the two 3’ flaps containing the edited sequence anneal with ...transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration with the Vallabh/Minikel Lab, the Weissman Lab...

...manuscript review/submission process at JEI. Chapter 3 - JEI target audience and their current readership...working in a lab for the first time, and thinking about the future. Many thanks to our guests,Olivia Ho-Shing...resources Use our Plasmids 101 eBook to teach students about basic plasmid biology Use or CRISPR 101 eBook to...

...help ensure ensure equal distribution. Lab tip #3: Barcode everything Use barcodes and scanners to keep...locate materials quickly and easily. Learn more about using barcodes. Check out our “Lab Tip” filter...

...a truncated version of the original Tol2 and has ~3-fold increase in transposition activity  Tol2 transposon...the QCascade DNA-targeting module with the crRNA, 3) a transposase complex made up by three transposase... up to 10 kB in size in bacteria.   Figure 3: INTEGRATE system for RNA guided transposon insertion...Addgene blog Learn about the INTEGRATE system for bacterial genome engineering  Read about the Sleeping Beauty...DNA transposons are most commonly used in the laboratory for genome manipulation. When transposons are...Applications of transposons Now that you’ve learned about some of the popular transposon systems, let’s take...2020) Jump around: transposons in and out of the laboratory. F1000Res 9:135. https://doi.org/10.12688/f1000research...

...pulse of 500 nm light (turn on psRaf1) followed by 3 seconds of 400 nm light (turn off psRaf1). After this...is responsible for this MEK1/2 negative feedback. 3. Studying signaling pathways in vivo Expressing photoswitchable...Additional Resources on the Addgene Blog Learn More about Photoactivatable Fluorescent Proteins Control Protein...

...produce yellow, cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created...Find the plasmids from this publication at Addgene. 3. Ma, Hanhui, et al. “Multiplexed labeling of genomic...Additional Resources on the Addgene Blog Read More about Fluorescent Protein-Based Tools Learn Some of the...

...proposal successful. Typical proposals consist of 3 to 5 aims, with secondary milestones to meet along...application to see how funds were allocated. Step 3: Get feedback from your peers and advisors Once a...first. Additionally, reach out to peers and collaborators who have succeeded at grant writing to see what...

...the stricter sequence requirement at the terminal 3’ PAM site (Fu et al. 2013; Pattanayak et al. 2013)...every CRISPR experiment. Off-target events due to >3 bp mismatches or that are sequence-independent are...PMID: 25566793. PubMed Central PMCID: PMC4349094. 3. Bell, Charles C., et al. "A high-throughput screening...et al. 2015). Meanwhile, Sanger sequencing is laborious, time consuming and cannot be applied to heterogenous...viable options for multiplex sequencing in many laboratories. Nevertheless, unbiased detection isn’t as sensitive...unbiased sequencing may not be worth the cost and labor when single clones are selected. Conversely, translational... Hough is a Biochemistry PhD candidate in the laboratory of Prof Steve Jackson at the Gurdon Institute...

... perspective on the field. 3. Highlight your network and your collaborators It’s a good idea to move around...before submitting - handing them your application 3 days before the deadline isn’t going to be much use...introduction to my area, a line about the research I will be doing, and a line about what I hope to discover....themselves many times and will have useful advice about what reviewers in your area are looking for. Remember...One way to add originality to your work is to collaborate with specialists in different fields that can...outcomes can include poster presentations, talks about your work, or even the successful use of your preliminary...quality application. Even following this advice, only about 50% of my grant applications have been successful...

... to show people where the lab magic is happening. 3) Volunteer – Step into science education Do you aim...-to-day business with popular work experiences of 3-10 days. And the Society of Experimental Biology offers...Sainsbury Laboratory. She is passionate about science communications and loves talking about science, ...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...break into science communication? Everybody goes about this in their own way. The career paths into science...Society gives training in media skills, writing about your research and residential communication. If ...

...materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various steps ...doi: 10.1038/nprot.2011.413. PubMed PMID: 22157972. 3. Britt JP, et al. Use of channelrhodopsin for activation...distance so ideal implant placement should be about 0.4-0.5mm above the site of your stimulation. The scatter...scatters a great deal once it enters tissue, placement above the site of stimulation guarantees that the majority...instead). Laser stimulation         In the video above, Channelrhodopsin 2 (Chr2) has been expressed in...These lasers are relatively cheap and will run you about $2-3,000/laser. However, an important caveat is ...output of the laser must be controlled. As discussed above, each opsin variant has slightly different activation...

... to have either salaries raised or hours tracked, 3% of postdocs were at institutions focused only on ...Previous Article: Changing Labor Laws Bring Increased Postdoc Wages Read about the Advocating for Science... of $47,476 per year, under updates to the Fair Labor Standards Act (FLSA). This was due to the threshold...postdocs - and in some cases, faculty - have found out about what their institution is doing from Future of Research...Science Symposium and Workshop Learn about a Variety of Science Careers Resources on Addgene.org Find Career...

...PMID: 27175853. Pubmed Central PMCID: PMC4866822. 3. McDowell, Gary S., et al. "Shaping the Future of ...perspective from junior scientists."F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central PMCID...purposes of labor laws, rather than as trainees in a special category, as has been the ‘labor-of-love’ philosophy...passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which mandates that all workers...event of note is a ruling in August by the National Labor Relations Board to allow graduate students who work...

...expression (Sapozhnikov and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation...work well, and up to 10 base pairs away (especially 3’ of PAM) might work too.  High dCas9 and gRNA expression...Daniel M. Sapozhnikov, a graduate student in the laboratory of Dr. Moshe Szyf at McGill University. Some ...expression, confounding any potential conclusions about the role of methylation (Hrit et al. 2018, Kaas ...Daniel M. Sapozhnikov is a graduate student in the laboratory of Dr. Moshe Szyf at McGill University, working...

...position at the Genome Institute of Singapore. Chapter 3: Biology research and resources in Singapore. Chapter...getting a lab up and running, and learn a little bit about what it was like for Wei Leong to work in George...